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Genemate blue ultra autoradiography film

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Genemate Blue ultra-autoradiography film is a high-performance X-ray film designed for autoradiographic applications. It is a photographic film that is used to detect and visualize radioactive signals in various scientific experiments, such as DNA sequencing, protein analysis, and gene expression studies.

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Lab products found in correlation

Np 40 buffer, by Thermo Fisher Scientific (5 mentions) Nitrocellulose membrane, by Bio-Rad (5 mentions) Bis tris gel, by Thermo Fisher Scientific (5 mentions) Dc protein assay kit, by Bio-Rad (4 mentions) Dry milk, by Merck Group (2 mentions) Complete protease inhibitor cocktail, by Roche (2 mentions) Phosstop phosphatase inhibitor cocktail, by Roche (2 mentions) Nupage sample reducing agent, by Thermo Fisher Scientific (2 mentions) Nupage lds sample buffer, by Thermo Fisher Scientific (2 mentions) Complete protease inhibitor, by Roche (2 mentions) Phosphatase inhibitor, by Roche (2 mentions) Cocktail of phosphatase inhibitors, by Roche (1 mentions) Lds sample buffer, by Thermo Fisher Scientific (1 mentions) Complete protease inhibitor tablet, by Roche (1 mentions) Dc protein assay, by Bio-Rad (1 mentions)

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Autoradiography film

5 protocols using genemate blue ultra autoradiography film

1

Western Blot Analysis of Protein Expression

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Cells were collected by trypsinization and lysed at 4°C in NP40 buffer (Invitrogen) supplemented with complete protease inhibitor cocktail (Roche), PhosSTOP phosphatase inhibitor cocktail (Roche) and PMSF (1 mM). Protein concentrations were determined with the Biorad DC protein assay kit (Bio-Rad). Whole-cell protein lysates were resolved on 4%–12% Bis-Tris gels (Invitrogen) and transferred to nitrocellulose membranes (Bio-Rad). After blocking nonspecific binding sites for 1 h using 5% dry milk (Sigma) in Tris-buffered saline (TBS) supplemented with 0.2% Tween-20 (TBS-T), membranes were incubated overnight with primary antibody at 4°C. Chemiluminescent detection was performed with the appropriate secondary antibodies and developed using Genemate Blue ultra autoradiography film (VWR).
Gao Y., Volegova M., Nasholm N., Das S., Kwiatkowski N., Abraham B.J., Zhang T., Gray N.S., Gustafson C., Krajewska M, & George R.E. (2022). Synergistic Anti-Tumor Effect of Combining Selective CDK7 and BRD4 Inhibition in Neuroblastoma. Frontiers in Oncology, 11, 773186.
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2

Western Blot Protein Analysis

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Cell lysates were prepared in NP-40 buffer (Invitrogen) containing complete protease inhibitor (Roche), phosphatase inhibitor (Roche) and phenylmethylsulfonyl fluoride (PMSF). Lysates were incubated on ice for 20 min and centrifuged at 16,000 g for 15 min at 4°C. Protein concentrations of the supernatants were determined with the DC protein assay kit (Bio-Rad). Protein samples were denatured using NuPAGE LDS sample buffer (Invitrogen) and NuPAGE sample reducing agent (Invitrogen). Proteins were then separated on precast 4%–12% Bis-Tris gels (Invitrogen) and transferred to nitrocellulose membranes (Bio-Rad). After blocking in blocking buffer (5% dry milk in TBS with 0.2% Tween-20), membranes were incubated with the primary antibody overnight at 4°C, washed three times with TBST buffer, chemiluminescent detection performed with the appropriate secondary antibodies and developed using Genemate Blue ultra-autoradiography film (VWR).
Huang H., Gont A., Kee L., Dries R., Pfeifer K., Sharma B., Debruyne D.N., Harlow M., Sengupta S., Guan J., Yeung C.M., Wang W., Hallberg B., Palmer R.H., Irwin M.S, & George R.E. (2021). Extracellular domain shedding of the ALK receptor mediates neuroblastoma cell migration. Cell reports, 36(2), 109363.
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3

Whole Cell Protein Extraction and Western Blotting

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Cells were collected by trypsinization and lysed at 4 °C in NP40 buffer (Invitrogen) supplemented with complete protease inhibitor cocktail (Roche), PhosSTOP phosphatase inhibitor cocktail (Roche) and PMSF (1 mM). Protein concentrations were determined with the Biorad DC protein assay kit (Bio-Rad). Whole cell protein lysates were resolved on 4–12% Bis-Tris gels (Invitrogen) and transferred to nitrocellulose membranes (Bio-Rad). After blocking nonspecific binding sites for 1 h using 5% dry milk (Sigma) in Tris-buffered saline (TBS) supplemented with 0.2% Tween-20 (TBS-T), membranes were incubated overnight with primary antibody at 4 °C. Chemiluminescent detection was performed with the appropriate secondary antibodies and developed using Genemate Blue ultra-autoradiography film (VWR). Uncropped versions of all western blots can be found in Supplementary Fig. 9.
Krajewska M., Dries R., Grassetti A.V., Dust S., Gao Y., Huang H., Sharma B., Day D.S., Kwiatkowski N., Pomaville M., Dodd O., Chipumuro E., Zhang T., Greenleaf A.L., Yuan G.C., Gray N.S., Young R.A., Geyer M., Gerber S.A, & George R.E. (2019). CDK12 loss in cancer cells affects DNA damage response genes through premature cleavage and polyadenylation. Nature Communications, 10, 1757.
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4

Western Blot Analysis of Protein Targets

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Cell or tumour tissue was lysed in NP-40 buffer (Invitrogen) containing a 1× complete protease inhibitor tablet (Roche) per 10 ml buffer and a cocktail of phosphatase inhibitors (Roche). Protein concentration was measured using the DC Protein Assay (Bio-Rad); protein (50 μg) was denatured in LDS sample buffer with reducing agent (Invitrogen), separated on precast 4–12% Bis-Tris gels (Life Technologies) and transferred to nitrocellulose membranes (Bio-Rad). Membranes were incubated in blocking buffer (5% dry milk in TBS with 0.2% Tween-20) for 1 h, and then incubated in the primary antibody in blocking buffer overnight at 4 °C. Chemiluminescent detection was performed with the appropriate secondary antibodies and developed using Genemate Blue ultra-autoradiography film (VWR). The actin loading controls for the protein samples shown in the immunoblots of the following panels (two independent mouse tumour samples, and cell lines representative of two independent experiments) are the same because the samples were run on a single gel but probed for pALK, ALK (Extended Data Fig. 2a), MYCN (Extended Data Fig. 3e) and BORIS (Extended Data Fig. 4a), respectively.
Debruyne D.N., Dries R., Sengupta S., Seruggia D., Gao Y., Sharma B., Huang H., Moreau L., McLane M., Day D.S., Marco E., Chen T., Gray N.S., Wong K.K., Orkin S.H., Yuan G.C., Young R.A, & George R.E. (2019). BORIS promotes chromatin regulatory interactions in treatment-resistant cancer cells. Nature, 572(7771), 676-680.
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5

Western Blot Protein Analysis

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Cell lysates were prepared in NP-40 buffer (Invitrogen) containing complete protease inhibitor (Roche), phosphatase inhibitor (Roche) and phenylmethylsulfonyl fluoride (PMSF). Lysates were incubated on ice for 20 min and centrifuged at 16,000 g for 15 min at 4°C. Protein concentrations of the supernatants were determined with the DC protein assay kit (Bio-Rad). Protein samples were denatured using NuPAGE LDS sample buffer (Invitrogen) and NuPAGE sample reducing agent (Invitrogen). Proteins were then separated on precast 4%–12% Bis-Tris gels (Invitrogen) and transferred to nitrocellulose membranes (Bio-Rad). After blocking in blocking buffer (5% dry milk in TBS with 0.2% Tween-20), membranes were incubated with the primary antibody overnight at 4°C, washed three times with TBST buffer, chemiluminescent detection performed with the appropriate secondary antibodies and developed using Genemate Blue ultra-autoradiography film (VWR).
Huang H., Gont A., Kee L., Dries R., Pfeifer K., Sharma B., Debruyne D.N., Harlow M., Sengupta S., Guan J., Yeung C.M., Wang W., Hallberg B., Palmer R.H., Irwin M.S, & George R.E. (2021). Extracellular domain shedding of the ALK receptor mediates neuroblastoma cell migration. Cell reports, 36(2), 109363.
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